Sole Carbon Substrate Research Articles

Microbial Induced Biotechnological Processes for Biofuel Production from Waste Organics Conversion

In the current era there are huge quantities of waste organic matter available, creating a big burden to the environment. To address these issues, researchers started to apply effective and microbial induced biotechnological processes that can mitigate these waste matters. In this context, different nature of microbial systems are involved in hydrolysing the waste organic material into fermentable sugar. These can be easily consumed by specific microbial systems like Saccharomyces cerevisiae MTCC 3821 and Clostridium acetobutylicum that produced bioethanol and biobutanol, respectively. Saccharomyces cerevisiae was cultured in specific media and incubated at rotary shaker with 150 rpm at 30°C for 72 to 96 hours. Ethanol concentrations from different waste matters were found in the range of 1.2-1.5 g.L-1. Ethanol synthesis was done by shake flask experiment with addition of glucose (50 g.L-1) to waste organic hydrolyzed solution. Non-glucose media produced less than 3 g.L-1 ethanol but glucose media produced 4.5 g.L-1. Next, Clostridium acetobutylicum was grown in culture media containing waste organics as sole carbon substrate with pH 7 and then was incubated in anaerobic conditions at 35°C for 72 hours, produced butanol (0.7 to 1.25 g.L-1). This research work promoted biofuels synthesis by keeping a waste mitigation strategy.

Synthesis and Properties of Polyhydroxyalkanoates on Waste Fish Oil from the Production of Canned Sprats

The waste fish oil obtained from Baltic sprat waste in the production of canned sprats was studied as a sole carbon substrate for PHA synthesis by the wild-type strain Cupriavidus necator B-10646. Sprat oil contained a set of fatty acids with a chain length from C14 to C24, saturation factor 0.63, and provided bacterial growth and PHA synthesis. Bacteria metabolized fatty acids unevenly utilizing polyenoic acids and not using monoenoic and saturated acids. The bacterial biomass yield and the intracellular polymer concentration were 6.5 ± 0.5 g/L and 65 ± 5% by fed-batch culture in flasks. The synthesized PHAs were three-component copolymers with a predominance (97–98 mol.%) of 3-hydroxybutyrate monomers and small inclusions of 3-hydroxyvalerate and 3-hydroxyhexanoate; the ratio of monomers changed slightly depending on the sprat oil concentration. The series of samples had a temperature (Tmelt) of 158–165 °C, a molecular weight (Mw) of 540–760 kDa, and a degree of crystallinity (Cx) of 66–72%. For the first time, the waste fish oil from the production of sprats studied as a carbon substrate is a promising, affordable, and renewable substrate for PHA biosynthesis.

Statistical optimization of P(3HB-co-3HHx) copolymers production by Cupriavidus necator PHB−4/pBBR_CnPro-phaCRp and its properties characterization

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] is a bacterial copolymer in the polyhydroxyalkanoates (PHAs) family, a next-generation bioplastic. Our research team recently engineered a newly P(3HB-co-3HHx)-producing bacterial strain, Cupriavidus necator PHB−4/pBBR_CnPro-phaCRp. This strain can produce P(3HB-co-2 mol% 3HHx) using crude palm kernel oil (CPKO) as a sole carbon substrate. However, the improvement of P(3HB-co-3HHx) copolymer production by this strain has not been studied so far. Thus, this study aims to enhance the production of P(3HB-co-3HHx) copolymers containing higher 3HHx monomer compositions using response surface methodology (RSM). Three significant factors for P(3HB-co-3HHx) copolymers production, i.e., CPKO concentration, sodium hexanoate concentration, and cultivation time, were studied in the flask scale. As a result, a maximum of 3.6 ± 0.4 g/L of P(3HB-co-3HHx) with 4 mol% 3HHx compositions was obtained using the RSM optimized condition. Likewise, the higher 3HHx monomer composition (5 mol%) was obtained when scaling up the fermentation in a 10L-stirrer bioreactor. Furthermore, the produced polymer's properties were similar to marketable P(3HB-co-3HHx), making this polymer suitable for a wide range of applications.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma.

Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Evaluating the soil microbe community‐level physiological profile using EcoPlate and soil properties at 33 forest sites across Japan

AbstractWe present the largest freely available EcoPlate dataset for Japan, comprising data collected from a network of 33 natural forest sites (77 plots) in regions of East Asia ranging from cool temperate to subtropical. EcoPlate is a 96‐well microplate that contains three repeated sets of 31 response wells with different sole carbon substrates. The utilization of each carbon substrate by the microbial community is quantified by the color density of the well during incubation. EcoPlate can provide a multifunctional index of a soil microbial community. Soil properties (water content, carbon [C] and nitrogen [N] contents, the C/N ratio, and pH) that are essential for interpreting the EcoPlate results were also measured. The network is part of the Forest and Grassland Survey of the Monitoring Sites 1000 Project established by the Ministry of the Environment. Using a standardized protocol, soil was sampled between September and December 2020. A preliminary principal component analysis was performed on the temporal integration of color density using 31 substrates. For the temporal integration, we calculated the cumulative amount of color development by integrating the color density development curve. PC1 contributed 55.6% and is thought to represent the magnitude of the overall absorbance of all substrates. The model of environmental factors including elevation and the model of soil properties, including water content and pH, were selected as the best‐fit models for variation in PC1. EcoPlate data enable the meta‐analysis of comparative studies among forest types and testing community ecology and ecosystem function hypotheses at broad spatial scales. The complete data set for this abstract published in the Data Paper section of the journal is available in electronic format in MetaCat in JaLTER at http://db.cger.nies.go.jp/JaLTER/metacat/metacat/ERDP‐2022‐01.1/jalter‐en.

A fermentation process for the production of poly(3-hydroxybutyrate) using waste cooking oil or waste fish oil as inexpensive carbon substrate

The utilization of waste cooking oil (WCO) or waste fish oil (WFO) as inexpensive carbon substrate for the production of poly(3-hydroxybutyrate) (PHB) by Cupriavidus necator H16 was investigated. Fed-batch cultivation mode in bioreactor was applied in this study. High cell dry weight (CDW) of 135.1 g/L, PHB content of 76.9 wt%, PHB productivity of 1.73 g/L/h, and PHB yield of 0.8 g/g were obtained from WCO. In the case of WFO, the CDW, PHB content, PHB productivity, and PHB yield were 114.8 g/L, 72.5 wt%, 1.73 g/L/h, and 0.92 g/g, respectively. The PHB productivity and yield obtained in the current study from WCO or WFO are among the highest reported so far for PHA production using oils as sole carbon substrate, suggesting that both WCO and WFO can be used as inexpensive carbon substrates for the production of PHA on an industrial scale.

Effect of macronutrients and of anaerobic digestate on the heterotrophic cultivation of Chlorella vulgaris grown with glycerol.

The aim of this work was to investigate the kinetics of the heterotrophic growth of Chlorella vulgaris as a means of producing bio-oil for biodiesel production. Glycerol was used as the sole organic carbon substrate. Anaerobic digestate from a local plant was used to examine its effect on the kinetics and the protein and lipid content of the biomass. The effect of the initial carbon and nitrogen concentrations on the carbon uptake rate was studied independently. In the one set of five experiments, the organic carbon in the form of glycerol varied from 0.27 to 5.36g L-1, while the concentration of atomic nitrogen was held constant and equal to 45.4mg L-1. The Co/No ratio varied from 6 to 118.1. In the second set, also of five experiments, the organic carbon was held constant and equal to 3.3g L-1 and atomic nitrogen varied from 22.7 to 450mg L-1. The Co/No ratio varied from 7.3 to 145.4. In the third set of experiments, anaerobic digestate was added in increasing amounts into the culture media from 4 to 16%. It was found that the carbon uptake rate as well as the lipid and protein content depended on the Co/No ratio. Increasing ratios of Co/No led to higher carbon uptake rates, higher lipid content, and lower protein content. The initial nitrogen concentration was also found to affect the growth rate of C. vulgaris. The addition of anaerobic digestate did not affect appreciably the protein and lipid content of the biomass, while the addition of anaerobic digestate up to 16% in the culture medium increased the carbon uptake rate by about 24%.

Utilization of agro-industrial waste for biosurfactant production under submerged fermentation and its synergistic application in biosorption of Pb2+

The biosurfactant production process was optimized using raw orange peel extract (ST reduction = 33.04 dyne cm−1; biosurfactant yield = ~3.7 g L−1) and the extracted metabolite was characterized in terms of its nature, and class/family. To the best of our knowledge, a first report utilizing biowaste as a sole carbon substrate for simultaneous biosurfactant production and Pb2+ removal under submerged fermentation by Bacillus haynesii strain E1. The results depicted the extracted biosurfactant to be of lipopeptide nature belonging to surfactin family having 50 mg L−1 of CMC. The crude biosurfactant was found to be tensioactive at temperature 70 °C, 6% salt concentration, and varying pH range. The biosurfactant-producing bacterium effectively remediated Pb2+ (high MIC = 2200 mg L−1) with a maximum adsorption capacity of 196.08 mg g−1. The biosorption mechanism followed Langmuir (R2 = 0.9724) and Pseudo second-order adsorption kinetics (R2 = 0.9996; 200 mg L−1).

Sustainable biosynthesis of chemicals from methane and glycerol via reconstruction of multi-carbon utilizing pathway in obligate methanotrophic bacteria.

SummaryObligate methanotrophic bacteria can utilize methane, an inexpensive carbon feedstock, as a sole energy and carbon substrate, thus are considered as the only nature‐provided biocatalyst for sustainable biomanufacturing of fuels and chemicals from methane. To address the limitation of native C1 metabolism of obligate type I methanotrophs, we proposed a novel platform strain that can utilize methane and multi‐carbon substrates, such as glycerol, simultaneously to boost growth rates and chemical production in Methylotuvimicrobium alcaliphilum 20Z. To demonstrate the uses of this concept, we reconstructed a 2,3‐butanediol biosynthetic pathway and achieved a fourfold higher titer of 2,3‐butanediol production by co‐utilizing methane and glycerol compared with that of methanotrophic growth. In addition, we reported the creation of a methanotrophic biocatalyst for one‐step bioconversion of methane to methanol in which glycerol was used for cell growth, and methane was mainly used for methanol production. After the deletion of genes encoding methanol dehydrogenase (MDH), 11.6 mM methanol was obtained after 72 h using living cells in the absence of any chemical inhibitors of MDH and exogenous NADH source. A further improvement of this bioconversion was attained by using resting cells with a significantly increased titre of 76 mM methanol after 3.5 h with the supply of 40 mM formate. The work presented here provides a novel framework for a variety of approaches in methane‐based biomanufacturing.

Methylobacterium ajmalii sp. nov., Isolated From the International Space Station.

Four strains belonging to the family of Methylobacteriaceae were isolated from different locations on the International Space Station (ISS) across two consecutive flights. Of these, three were identified as Gram-negative, rod-shaped, catalase-positive, oxidase-positive, motile bacteria, designated as IF7SW-B2T, IIF1SW-B5, and IIF4SW-B5, whereas the fourth was identified as Methylorubrum rhodesianum. The sequence similarity of these three ISS strains, designated as IF7SW-B2T, IIF1SW-B5, and IIF4SW-B5, was <99.4% for 16S rRNA genes and <97.3% for gyrB gene, with the closest being Methylobacterium indicum SE2.11T. Furthermore, the multi-locus sequence analysis placed these three ISS strains in the same clade of M. indicum. The average nucleotide identity (ANI) values of these three ISS strains were <93% and digital DNA-DNA hybridization (dDDH) values were <46.4% with any described Methylobacterium species. Based on the ANI and dDDH analyses, these three ISS strains were considered as novel species belonging to the genus Methylobacterium. The three ISS strains showed 100% ANI similarity and dDDH values with each other, indicating that these three ISS strains, isolated during various flights and from different locations, belong to the same species. These three ISS strains were found to grow optimally at temperatures from 25 to 30°C, pH 6.0 to 8.0, and NaCl 0 to 1%. Phenotypically, these three ISS strains resemble M. aquaticum and M. terrae since they assimilate similar sugars as sole carbon substrate when compared to other Methylobacterium species. Fatty acid analysis showed that the major fatty acid produced by the ISS strains are C18:1−ω7c and C18:1−ω6c. The predominant quinone was ubiquinone 10, and the major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, and an unidentified lipid. Therefore, based on genomic, phylogenetic, biochemical, and fatty acid analyses, strains IF7SW-B2T, IIF1SW-B5, and IIF4SW-B5, are assigned to a novel species within the genus Methylobacterium, and the name Methylobacterium ajmalii sp. nov. is proposed. The type strain is IF7SW-B2T (NRRL B-65601T and LMG 32165T).

Nitrogen application and intercropping change microbial community diversity and physicochemical characteristics in mulberry and alfalfa rhizosphere soil

Intercropping of mulberry (Morus alba L.) and alfalfa (Medicago sativa L.) is a new forestry-grass compound model in China, which can provide high forage yields with high protein. Nitrogen application is one of the important factors determining the production and quality of this system. To elucidate the advantages of intercropping and nitrogen application, we analyzed the changes of physicochemical properties, enzyme activities, and microbial communities in the rhizosphere soil. We used principal components analysis (PCA) and redundancy discriminators analysis to clarify the relationships among treatments and between treatments and environmental factors, respectively. The results showed that nitrogen application significantly increased pH value, available nitrogen content, soil water content (SWC), and urea (URE) activity in rhizosphere soil of monoculture mulberry. In contrast, intercropping and intercropping + N significantly decreased pH and SWC in mulberry treatments. Nitrogen, intercropping and intercropping + N sharply reduced soil organic matter content and SWC in alfalfa treatments. Nitrogen, intercropping, and intercropping + N increased the values of McIntosh diversity (U), Simpson diversity (D), and Shannon–Weaver diversity (H′) in mulberry treatments. However, PCA scatter plots showed clustering of monoculture mulberry with nitrogen (MNE) and intercropping mulberry without nitrogen (M0). Intercropping reduced both H′ and D but nitrogen application showed no effect on diversity of microbial communities in alfalfa. There were obvious differences in using the six types of carbon sources between mulberry and alfalfa treatments. Nitrogen and intercropping increased the numbers of sole carbon substrate in mulberry treatments where the relative use rate exceeded 4%. While the numbers declined in alfalfa with nitrogen and intercropping. RDA indicated that URE was positive when intercropping mulberry was treated with nitrogen, but was negative in monoculture alfalfa treated with nitrogen. Soil pH and SWC were positive with mulberry treatments but were negative with alfalfa treatments. Intercropping with alfalfa benefited mulberry in the absence of nitrogen application. Intercropping with alfalfa and nitrogen application could improve the microbial community function and diversity in rhizosphere soil of mulberry. The microbial community in rhizosphere soil of mulberry and alfalfa is strategically complementary in terms of using carbon sources.

Enhancement of polyhydroxyalkanoate production by co-feeding lignin derivatives with glycerol in Pseudomonas putida KT2440

BackgroundEfficient utilization of all available carbons from lignocellulosic biomass is critical for economic efficiency of a bioconversion process to produce renewable bioproducts. However, the metabolic responses that enable Pseudomonas putida to utilize mixed carbon sources to generate reducing power and polyhydroxyalkanoate (PHA) remain unclear. Previous research has mainly focused on different fermentation strategies, including the sequential feeding of xylose as the growth stage substrate and octanoic acid as the PHA-producing substrate, feeding glycerol as the sole carbon substrate, and co-feeding of lignin and glucose. This study developed a new strategy—co-feeding glycerol and lignin derivatives such as benzoate, vanillin, and vanillic acid in Pseudomonas putida KT2440—for the first time, which simultaneously improved both cell biomass and PHA production.ResultsCo-feeding lignin derivatives (i.e. benzoate, vanillin, and vanillic acid) and glycerol to P. putida KT2440 was shown for the first time to simultaneously increase cell dry weight (CDW) by 9.4–16.1% and PHA content by 29.0–63.2%, respectively, compared with feeding glycerol alone. GC–MS results revealed that the addition of lignin derivatives to glycerol decreased the distribution of long-chain monomers (C10 and C12) by 0.4–4.4% and increased the distribution of short-chain monomers (C6 and C8) by 0.8–3.5%. The 1H–13C HMBC, 1H–13C HSQC, and 1H–1H COSY NMR analysis confirmed that the PHA monomers (C6–C14) were produced when glycerol was fed to the bacteria alone or together with lignin derivatives. Moreover, investigation of the glycerol/benzoate/nitrogen ratios showed that benzoate acted as an independent factor in PHA synthesis. Furthermore, 1H, 13C and 31P NMR metabolite analysis and mass spectrometry-based quantitative proteomics measurements suggested that the addition of benzoate stimulated oxidative-stress responses, enhanced glycerol consumption, and altered the intracellular NAD+/NADH and NADPH/NADP+ ratios by up-regulating the proteins involved in energy generation and storage processes, including the Entner–Doudoroff (ED) pathway, the reductive TCA route, trehalose degradation, fatty acid β-oxidation, and PHA biosynthesis.ConclusionsThis work demonstrated an effective co-carbon feeding strategy to improve PHA content/yield and convert lignin derivatives into value-added products in P. putida KT2440. Co-feeding lignin break-down products with other carbon sources, such as glycerol, has been demonstrated as an efficient way to utilize biomass to increase PHA production in P. putida KT2440. Moreover, the involvement of aromatic degradation favours further lignin utilization, and the combination of proteomics and metabolomics with NMR sheds light on the metabolic and regulatory mechanisms for cellular redox balance and potential genetic targets for a higher biomass carbon conversion efficiency.

Regulatory interventions improve the biosynthesis of limiting amino acids from methanol carbon to improve synthetic methylotrophy in Escherichia coli.

Synthetic methylotrophy aims to engineer methane and methanol utilization pathways in platform hosts like Escherichia coli for industrial bioprocessing of natural gas and biogas. While recent attempts to engineer synthetic methylotrophs have proved successful, autonomous methylotrophy, that is, the ability to utilize methane or methanol as sole carbon and energy substrates, has not yet been realized. Here, we address an important limitation of autonomous methylotrophy in E. coli: the inability of the organism to synthesize several amino acids when grown on methanol. We targeted global and local amino acid regulatory networks. Those include removal of amino acid allosteric feedback inhibition (argAH15Y , ilvAL447F , hisGE271K , leuAG462D , proBD107N , thrAS345F , trpES40F ), knockouts of transcriptional repressors (ihfA, metJ); and overexpression of amino acid biosynthetic operons (hisGDCBHAFI, leuABCD, thrABC, trpEDCBA) and transcriptional regulators (crp, purR). Compared to the parent methylotrophic E. coli strain that was unable to synthesize these amino acids from methanol carbon, these strategies resulted in improved biosynthesis of limiting proteinogenic amino acids (histidine, leucine, lysine, methionine, phenylalanine, threonine, tyrosine) from methanol carbon. In several cases, improved amino acid biosynthesis from methanol carbon led to improvements in methylotrophic growth in methanol minimal medium supplemented with a small amount of yeast extract. This study addresses a key limitation currently preventing autonomous methylotrophy in E. coli and possibly other synthetic methylotrophs and provides insight as to how this limitation can be alleviated via global and local regulatory modifications.

Effects of Several Physicochemical Factors on the Biodegradation of 1,4-Dioxane by Microbial Consortium CDIK-3 Enriched from Industrial Wastewater

Objective : With the ultimate aim of application for effluent treatment, the relationships between 1,4-dioxane degradation by microbial consortium CDIK-3 and several relevant physicochemical environmental parameters were investigated.Methods : Microbial consortium CDIK-3 enriched from an industrial wastewater sample developed to grow aerobically with 1,4-dioxane as the sole carbon substrate was used to evaluate the degradation of 1,4-dioxane depending on the various environmental parameters and HPLC methodology was used to measure residual 1,4-dioxane in the test consortium.Results and Discussion : The microbial consortium, designated CDIK-3, was originally enriched from industrial wastewater sample. Complete depletion of 1,000 mg/L 1,4-dioxane was achieved in this experiment within 168 h. Effective 1,4-dioxane degradation was observed in slightly alkaline conditions (pH 8) and 30℃. The effect of supplemented carbon and nitrogen (&lt;i&gt;e.g&lt;/i&gt;., glucose, yeast extract, ethylene glycol, tetrahydrofuran) on the 1,4-dioxane degradation by the CDIK-3 test culture was evaluated. The results indicated that the addition of glucose, yeast extract, or tetrahydrofuran accelerated bacterial growth compared to growth in the absence of supplemented carbons. In contrast, the addition of low concentrations (≤100 mg/L) of ethylene glycol accelerated 1,4-dioxane degradation, but the degradation of 1,4-dioxane was delayed in the culture to which high concentrations (≥200 mg/L) of ethylene glycol were added.Conclusions : 1,4-Dioxane as a wastewater contaminant can be removed by the microbial consortium CDIK-3 enriched from industrial wastewater. Effective 1,4-dioxane removal was obtained by the control of various physicochemical parameters. These results provide clues for understanding improved 1,4-dioxane degradation by microbial consortium CDIK-3 with relevant environmental parameters and possible applications in the processing of industrial wastewater containing relatively high 1,4-dioxane concentrations.

Triggering the stringent response enhances synthetic methanol utilization in Escherichia coli

Synthetic methylotrophy aims to engineer methane and methanol utilization pathways in platform hosts like Escherichia coli for industrial bioprocessing of natural gas and biogas. While recent attempts to engineer synthetic methylotrophs have proved successful, autonomous methylotrophy, i.e. the ability to utilize methane or methanol as sole carbon and energy substrates, has not yet been realized. Here, we address an important limitation of autonomous methylotrophy in E. coli: the inability of the organism to synthesize several amino acids when grown on methanol. By activating the stringent/stress response via ppGpp overproduction, or DksA and RpoS overexpression, we demonstrate improved biosynthesis of proteinogenic amino acids via endogenous upregulation of amino acid synthesis pathway genes. Thus, we were able to achieve biosynthesis of several limiting amino acids from methanol-derived carbon, in contrast to the control methylotrophic E. coli strain. This study addresses a key limitation currently preventing autonomous methylotrophy in E. coli and possibly other synthetic methylotrophs and provides insight as to how this limitation can be alleviated via stringent/stress response activation.

Batch degradation of trichloroethylene using oleaginous Rhodococcus opacus in a two-phase partitioning bioreactor and kinetic study

In the present study, we present the biodegradation of trichloroethylene (TCE) for the first time using pure bacterial strain of Rhodococcus opacus PD630. TCE utilized as the sole carbon substrate by R. opacus in single phase system and was reached to highest specific growth rate of 0.015 h−1 at the concentration of 75 mg L−1. To further enhance the degradation performance even at high concentration (1000 mg L−1), two-phase partitioning bioreactor (TPPB) was developed. Silicone oil (20%, v/v) was selected as non-aqueous phase solvent to achieve high metabolic activity of bacteria. Furthermore, the experimental results of TCE degradation and specific TCE utilization rate were fitted to various kinetic models. Our experimental results clearly revealed that TPPB system having R. opacus as biological source and silicone oil as solvent was highly efficient and successful for removing high concentrated TCE compared with single phase system.

Deciphering the role of dissolved oxygen and N-acetyl glucosamine in governing higher molecular weight hyaluronic acid synthesis in Streptococcus zooepidemicus cell factory.

The present study is focused on systematic process and kinetic investigation of hyaluronic acid (HA) production strategy unraveling the role of dissolved oxygen (DO) and N-acetyl glucosamine (GlcNAc) towards the enhancement of HA titer and its molecular weight. Maintaining excess DO levels (10-40% DO) through DO-stat control and the substitution of GlcNAc at a range (5-20g/L) with glucose (Glc) critically influenced HA production. DO-stat control strategy yielded a promising HA titer (2.4g/L) at 40% DO concentration. Controlling DO level at 20% (DO-stat) was observed to be optimum resulting in a significant HA production (2.1g/L) and its molecular weight ranging 0.98-1.45MDa with a consistent polydispersity index (PDI) (1.57-1.69). Substitution of GlcNAc with Glc at different proportions explicitly addressed the metabolic trade-off between HA titer and its molecular weight. GlcNAc substitution positively influenced the molecular weight of HA. The highest HA molecular weight (2.53MDa) of two-fold increase compared with glucose as sole carbon substrate and narrower PDI (1.35 ± 0.18) was achieved for the 10:20 (Glc:GlcNAc) proportion. A novice attempt on modeling the uptake of dual substrates (Glc and GlcNAc) by Streptococcus zooepidemicus for HA production was successfully accomplished using double Andrew's growth model and the kinetic parameters were estimated reliably.

A novel carbon monoxide fed moving bed biofilm reactor for sulfate rich wastewater treatment

In this study, a moving bed biofilm reactor was used for biodesulfuruization using CO as the sole carbon substrate. The effect of hydraulic retention time (HRT), sulfate loading rate and CO loading rate on sulfate and CO removal was examined. At 72, 48 and 24 h HRT, the sulfate removal was 93.5%, 91.9% and 80.1%, respectively. An increase in the sulfate loading reduced the sulfate reduction efficiency, which, however, was improved by increasing the CO flow rate into the MBBR. Best results in terms of sulfate reduction (>80%) were obtained for low inlet sulfate and high CO loading conditions. The CO utilization was very high at 85% throughout the study, except during the last phase of the continuous bioreactor operation it was around 70%. An artificial neural network based model was successfully developed and optimized to accurately predict the bioreactor performance in terms of both sulfate reduction and CO utilization. Overall, this study showed an excellent potential of the moving bed biofilm bioreactor for efficient sulfate reduction even under high loading conditions.

Ability of the So-Called Obligate Hydrocarbonoclastic Bacteria to Utilize Nonhydrocarbon Substrates Thus Enhancing Their Activities Despite their Misleading Name

BackgroundThe group of the so-called obligate hydrocarbonoclastic bacteria (OHCB) are marine microorganisms affiliated with the genera Alcanivorax, Cycloclasticus, Oleiphilus and Thalassolituus. This small group plays a major role in oil-bioremediation in marine ecosystems. Marinobacter and Planomicrobium are considered related to this group. The OHCB are claimed to be obligate to hydrocarbon nutrition. This study argues against this claim.ResultsFour Alcanivorax species, three Marinobacter species and Planomicrobium okeanokoites from the Arabian/Persian Gulf proved to be not obligate to hydrocarbon nutrition. Although the eight strains grew on crude oil, n-octadecane and phenanthrene as sole carbon substrates, their growth was weaker than on certain nonhydrocarbon, organic compounds viz. peptone, glutamic acid, pyruvic acid, sucrose, mannose and others. Glucose and lactose failed to support the growth of seven of the eight tested strains. Mannose was utilized by five and sucrose by six strains. The well-known intermediate metabolite; pyruvic acid was utilized by all the eight strains, and lactic acid by five strains. In batch cultures, all the tested species consumed higher proportions of peptone than of n-alkanes and phenanthrene. When peptone and crude oil were provided together into the medium, the OHCB started to consume peptone first, and the enriched bacterial populations consumed oil effectively. Added nonhydrocarbon substrates biostimulated oil-consumption by all OHCB species.ConclusionThe tested OHCB species are not obligate hydrocarbon-utilizers. This provides them with the merit of survival, should their marine ecosystems become oil- or hydrocarbon-free. The fact that conventional, organic substrates biostimulated hydrocarbon-consumption by the tested bacterial species confirms the facultative nature of those organisms and is interesting from the practical point of view.

Biohydrogen by Rhodobacter sphaeroides during photo-fermentation: Mixed vs. sole carbon sources enhance bacterial growth and H2 production

Mixed carbon sources fermentation by bacteria is a promising approach for biohydrogen (H2) production biotechnology. In the present study, growth and Н2 production by purple bacteria Rhodobacter sphaeroides MDC6521 during mixed carbon sources (succinate + acetate, succinate + malate, and malate + acetate) photo-fermentation was investigated. The growth rate of bacteria in mixed carbon sources containing medium was of ∼0.33 h−1 which was considerably higher (1.3–1.7-fold) compared with sole carbon substrate containing one. Moreover, the H2 production during photo-fermentation of succinate and acetate mixture was of ∼6.5 mmol H2 g−1 (dry weight of biomass) and significantly more (∼2–3-fold) than that with appropriate sole sources and higher (1.5-fold) than that with succinate and malate mixture. Probably, supplementation of the mixed carbon sources into bacterial culture alters the mode of metabolism, resulting in enhanced H2 production, thus they can be preferable compared to the sole carbon source. The changed FOF1-ATPase activity of membrane vesicles suggested its important role in the increase of Н2 production efficiency. The results showed that mixed carbon sources provide more H2 than the sole carbon substrates and succinate with acetate mixture is better than succinate with malate.